Estimation of postmortem interval using histological and oxidative biomarkers in human bone marrow.

In forensic medicine, estimating the postmortem interval (PMI) is of great importance for the timeline and the reconstruction of the events surrounding death. Bone marrow (BM) is one of the largest organs in the body, with good resistance to autolysis and contamination. Therefore, the present study aims to correlate different postmortem intervals and bone marrow antioxidant enzyme levels using an Enzyme-linked immunosorbent assay (ELISA). In addition, detection of the changes in the histological structure of human bone marrow in relation to the time passed since death. BM samples from 20 forensic autopsy cadavers were obtained from cases referred to the Department of Forensic Medicine in the Ministry of Justice, Dakahlia Governorate, processed for histopathological examination as well as estimation of reduced glutathione (GSH), glutathione peroxidase (GPX), and glutathione reductase (GRX) using ELISA. Results of ELISA analysis showed a significant decrease in the level of antioxidant enzymes with increasing PMI; regarding histopathological examination, from 6 to > 18 h PMI, the changes in morphology after death were gradual, progressive, and regular, indicating great value in PMI determination. Also, 18 h of PMI showed loss of cellular details, absence of fat cells, and necrosis of BM with the nucleus dispersed as eosinophilic debris. Estimation of antioxidant enzymes level in human bone marrow using ELISA and detection of the changes in the histological structure of human bone marrow in relation to time passed since the death, either separately or in combination, can be used to estimate PMI accurately.

Expression of HnRNP A1, ZEB1, and E-cadherin in Hepatocellular carcinoma and their impact on patients' prognosis and survival.

Background: Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies in Egypt. HCCs usually have a poor prognosis because of late diagnosis, aggressive metastasis, and early invasion. Heterogeneous ribonucleoproteins (HnRNPs) are nuclear proteins that play a variety of roles in telomere formation, DNA repair, cell signaling, and gene regulation. .: Zincfinger Eboxbinding homeoboxes (ZEBs) are transcription factors that have a consistent inverse correlation with Ecadherin in numerous types of cancer and associated with poor prognosis. Aim: This study aimed to verify the prognostic expression of HnRNP A1, ZEB1, and E-cadherin in HCC. Settings and Design: The retrospective study consisted of 54 formalin-fixed paraffin-embedded tissue blocks of hepatocellular carcinoma. Methods and Material: Immunohistochemical staining was performed using antibodies against HnRNP A1, ZEB1, and E-cadherin. The patients were followed at the Clinical Oncology Department from May 2018 to July 2021. Statistical Analysis: SPSS version 20 using the Chi-square test to compare data and the Kaplan-Meier plot for comparing survival. Results: HnRNP A1 high positivity was detected in 59.3% of the cases, whereas negative E-cadherin and ZEB 1 expression presented in 37% and 70.4% of the patients, respectively. A statistically significant relation was present between HnRNP A1, ZEB1, E-cadherin, and various clinicopathological variables. The mean progression-free survival and overall survival in low HnRNP A1 and negative ZEB1 expressions were longer than those exhibited in high HnRNP A1 and positive ZEB1 expressions. Conclusion: HnRNP A1 and ZEB1 expressions are poor prognostic factors of HCC. E-cadherin has an important role in the development of differentiated HCCs and favorable outcome.

Outcome of Patients With Head and Neck Squamous Cell Carcinoma Can be Predicted by Expression of eIF4E and Osteopontin in Free Surgical Margins.

Squamous cell carcinoma of the head and neck (HNSCC) is recognized as the third most common cause of death. Incomplete resection of the primary tumor is the main cause of local recurrence and poor prognosis in HNSCC. Histologic assessment in order to determine "tumor-free" margins could be inadequate because of malignant transformation occurs at the molecular level earlier than the morphologic level. The present study aimed to evaluate the prognostic significance of eukaryotic initiation factor 4E (eIF4E) and Osteopontin in the tumor cells and histologically tumor free surgical margins of HNSCC. This cohort study was performed on 60 cases of HNSCC diagnosed at the Department of Pathology and treated at the Clinical Oncology Department, Faculty of Medicine, Zagazig University. Our enrolled formalin fixed paraffin embedded biopsy specimens with their matched tumor free surgical margins from resected head and neck squamous cell carcinoma were immunostaind for eIF4E and Osteopontin markers. 65% of our HNSCC patients had eIF4 E positive cytoplasmic immunostaining and 70% of them exhibited Osteopontin staining. Two-thirds of the dead patients exhibited high Osteopontin positive staining, whereas the surviving group did not exhibit this high expression. Concerning eIF4E, 85% and 5% of the dead patients showed high and low eIF4E expression, respectively. Disease-free survival (DFS) and overall survival were significantly (P=0.000) different between high and negative expression of Osteopontin, high and negative expression of eIF4E. 84% of patients with eIF4E positive margins and 75% with Osteopontin positive margins had local recurrence. In addition, negative expression of eIF4E is associated with highly significant better DFS and overall survival (P=0.000 and 0.001), respectively, in the margin negative expression status, while negative expression of Osteopontin was significantly associated with better DFS but of no significance in overall survival outcome. Our findings suggest that tumor-free surgical margins in HNSCC may be redefined as histologically Osteopontin and eIF4E negative resection margins. However, multicenter prospective studies are required to further evaluate their clinical utility in the surgical management of primary HNSCC.

Protective roles of thymoquinone and vildagliptin in manganese-induced nephrotoxicity in adult albino rats.

Despite being important in the body's mechanisms, excessive accumulation of manganese (Mn) can induce severe toxicity in vital organs of the body. Thymoquinone (TQ) is extracted from Nigella sativa seeds which recently gained popularity as dietary supplements and plant-based antioxidants. Vildagliptin (VLD) is a dipeptidyl peptidase IV (DPPIV) inhibitor, approved as anti-hyperglycemic agents with cardioprotective and renoprotective effects. The present study aimed to investigate the nephrotoxicity of Mn and the potential protective effects of thymoquinone and vildagliptin. Sixty-four adult male albino rats were equally divided into 8 groups: group I (control, received no medication), group II (vehicle, received normal saline), group III (TQ, 50 mg/kg/day), group IV (VLD, 10 mg/kg/day), group V (MnCl2, 50 mg/kg/day), group VI (Mn+TQ), group VII (Mn+VLD), and group VIII (Mn+TQ+VLD). Groups VI, VII, and VIII, received the same previously mentioned doses. All drugs were orally gavaged for 12 weeks. Manganese administration resulted in an elevation in the levels of serum and tissues Mn, blood glucose, serum urea, creatinine, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and reduction in insulin, kidney superoxide dismutase (SOD), glutathione (GSH), and interleukin-10. Histopathological structural renal damage was detected associated with strong positive immunoexpression of caspase-3. On the other hand, individual or combined TQ and VLD administration with Mn significantly decreased the serum and tissue levels of Mn, declined the blood glucose, inflammatory markers, oxidative stress markers, ameliorated the histopathological effects, and down-regulated the immunoexpression of caspase-3. In conclusion, TQ and VLD co-administration elicited protective effects against Mn-induced nephrotoxicity.

SATB-1 and Her2 as predictive molecular and immunohistochemical markers for urothelial cell carcinoma of the bladder.

Studying bladder cancer molecular biology revealed the presence of genetic alterations. So, detection of molecular biomarkers that help in monitoring the disease, evaluating the prognosis of the patients, and their response to therapy is needed. In this study, we investigated the expression and the prognostic significance of SATB-1 and ERBB2 mRNA and protein by quantitative RT-PCR and immunohistochemical analysis in urothelial bladder cancer cases and the surrounding normal bladder tissue. The correlations between the expression of both markers and the clinicopathological parameters were performed with further analysis of the correlation between the expression of SATB-1 and ERBB2. Compared to control, the expression of SATB-1 and ERBB2 mRNA and protein in cancer tissues were significantly up-regulated (p< 0.05). Also, a positive correlation between both markers was found (r= 0.53, p< 0.001). Moreover, elevated levels of both markers were significantly associated with the stage, lymph node involvement at both mRNA and protein levels (p< 0.001). In conclusion, there is a clinical significance of SATB-1 and ERBB2 as potential biomarkers for predicting bladder cancer patients of aggressive behavior and poor prognosis.

Differential expression of NKX 3.1 and HOXB 13 in bone metastases originating from prostatic carcinoma among the Egyptian males.

BACKGROUND: The skeleton represents one of the most common sites to be affected by metastatic tumors. About 65 % of all bone metastases come from the breast cancer in females, and from the prostatic carcinoma in males. A probable diagnostic pitfall may be encountered during the process of decalcification of the bone metastases specimens. This study aimed to evaluate the diagnostic utility of NKX3.1 and HOXB13 and compare them with the traditionally used PSA for the detection of prostatic origin of bone metastases. MATERIAL AND METHODS: We analyzed 41 tissue specimens of bone metastases originating from prostatic carcinoma. Immunohistochemical staining of NKX3.1, HOXB13 and PSA was done with evaluation of their differential expression. RESULTS: NKX3.1, HOXB 13 and PSA were expressed in (41/41), (39/41) and (25/41) respectively of the analyzed cases. On comparing NKX3.1 and HOXB13 positive staining, there was a statistically significant difference (P = 0.000). In addition, the frequency of positive NKX3.1 expression in decalcified bone biopsies of metastatic prostatic adenocarcinoma is statistically higher than that of PSA immunostaining (P = 0.000). We found a statistically significant difference between HOXB13 and PSA positive immunostaining (P = 0.01). CONCLUSION: This study demonstrates that NKX3.1 and HOXB13 are more sensitive markers than PSA for the detection of prostate carcinoma metastatic to the bone following the decalcification process. We recommend use of NKX3.1 and HOXB13, rather than PSA, in the diagnosis of bone metastases originating from prostate cancer.

Diagnostic utility of amylase α-1A, MOC 31, and CD 82 in renal oncocytoma versus chromophobe renal cell carcinoma.

OBJECTIVE: Renal oncocytoma (RO) and chromophobe renal cell carcinoma (ChRCC) originate from the same cell origin, that is, the intercalated cells of the collecting duct.[1] In most cases, there are clear morphologic differences between RO and ChRCC; however, in some instances, overlapping features may be encountered and the differentiation between the two entities becomes difficult.[2] Several immunohistochemical markers with different expression patterns in ChRCC and RO have been described to rule out this dilemma. MATERIALS AND METHODS: About 47 primary renal neoplasms that had been diagnosed as RO or ChRCC were submitted for immunohistochemical staining of amylase α-1A (AMY1A), MOC 31, and CD 82. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy have been analyzed. RESULTS: AMY1A positivity was observed in all RO cases in our work with 91.7% sensitivity and 100% specificity in the diagnosis of RO. The PPV of its expression was (100%) and NPV (97.2%) with a diagnostic accuracy of 97.9%. A significant high expression of MOC 31 was observed in ChRCC compared to its expression in RO with a statistical significance (P < 0.001). In addition, we obtained 82.9% sensitivity and 91.7% specificity of MOC 31 expression in the diagnosis of ChRCC. The positive predictive value (PPV) was (96.7%), negative predictive value (NPV) (64.7%) with diagnostic accuracy (85.1%). In our studied cases, we detected positive immunoexpression of CD 82 in 10 cases (83.3%) of ChRCC. However, it was lost in all RO cases (100%). CD 82 sensitivity and specificity in differentiating ChRCC from RO were 100% and 83.3%, respectively. CONCLUSION: We propose MOC 31 and CD 82 as negative immunostains for RO, as these markers are commonly expressed in ChRCC. In conjunction with AMY1A strong immunopositivity in RO cases, we provide a triple panel of biomarkers (AMY1A, MOC 31, and CD 82) for the distinction between RO and ChRCC.

Mesenchymal Stem Cells: a Promising Therapeutic Tool for Acute Kidney Injury.

Acute kidney injury (AKI) is a rapid loss of renal function. It has high mortality rates. Still, renal replacement therapy is considered the best solution for recovering AKI. This opens a line of thought to develop an alternative therapy for it without complications. Mesenchymal stem cells are considered a new therapy for treating kidney diseases. The aim of this work was to address the anti-apoptotic, antioxidative, and pro-angiogenic effects of adipose tissue-derived MSCs (AD-MSCs) and bone marrow-MSCs (BM-MSCs) for treating AKI. Adult male Wistar rats were assigned into nine groups (n = 10): (1) the control group; (2) the AKI group, receiving cisplatin; (3) the AKI group treated with AD-MSCs (1 × 106); (4) the AKI group treated with AD-MSCs (2 × 106); (5) the AKI group treated with AD-MSCs (4 × 106); (6) the AKI group treated with losartan; (7) the AKI group treated with BM-MSCs (1 × 106); (8) the AKI group treated with BM-MSCs (2 × 106); and (9) the AKI group treated with BM-MSCs (4 × 106). The results showed a significant rise in creatinine, urea, and cystatin C (cys C) levels and upregulation of p38 mRNA, whereas a significant decline in NAD(P)H quinone oxidoreductase 1 (NQO-1) protein and downregulation of B-cell lymphoma-2 (Bcl-2) mRNA and vascular endothelial growth factor (VEGF) mRNA were recorded in AKI. MSCs could improve renal functions manifested by decreased urea, creatinine, and cys C levels; downregulation of p38; and upregulation of Bcl-2 and VEGF. Moreover, MSC therapy could induce NQO-1 in the treated rats relative to the untreated rats. So, cell-based therapy can reduce AKI through the antioxidative, anti-apoptotic, and pro-angiogenic properties of MSCs. Therefore, the findings received in this attempt create a fertile base for the setup of cell therapy in patients with AKI.

Alleviative effect of licorice on copper chloride-induced oxidative stress in the brain: biochemical, histopathological, immunohistochemical, and genotoxic study.

Although copper is an essential micronutrient involved in a variety of biological processes indispensable for sustaining life, it can be toxic when administered in excess. Licorice (Glycyrrhizaglabra) has been used in Chinese folk medicine for the treatment of various disorders. Licorice has the biological capabilities of detoxication, antioxidation, and antiinfection. Here, we test the hypothesis that licorice could ameliorate copper-induced neurotoxic and genotoxic effects in adult male albino rats. For this purpose, 48 adult male albino rats were randomized into five groups: group I (8 rats), untreated control; group II (16 rats), subdivided into; vehicle control IIa (8 rats) which received 1 mL saline twice weekly intraperitoneally for 8 weeks and vehicle control IIb (8 rats) received 0.5 mL distilled water/day orally gavaged for 8 weeks; group III (8 rats), treated with licorice dissolved in 0.5 mL of distilled water, 50 mg/kg b.w./day orally gavaged for 8 weeks; group IV (8 rats), copper chloride (CuCl2) dissolved in 0.5 mL saline, 7 mg/kg b.w. twice weekly intraperitoneal for 8 weeks; and group V (8 rats), CuCl2 + licorice (the same previously mentioned doses) licorice extract were orally given for 10 days before treatment was initiated then followed by CuCl2 intraperitoneally for 8 weeks. We found that CuCl2 exposure significantly increased brain oxidative stress as manifested by elevated malondialdehyde levels, decreased reduced glutathione content, and depressed antioxidant enzyme activities in brain tissues when compared with control groups. This was accompanied by histopathological changes in the form of increased cellularity and swelling of astrocytes that showed dense eosinophilic cytoplasm, pyknotic nuclei, and multiple apoptotic bodies that associated with degenerated neurons with deep eosinophilic cytoplasm. Also, strong Bax immunoreactions in the brain were detected. Furthermore, comet assay results confirmed CuCl2-related oxidative DNA damage. Notably, all these changes were partially ameliorated in rats treated concomitantly with licorice and CuCl2. Our results showed that licorice exerts protective effects against CuCl2-induced neuro- and genotoxicities. These effects may be attributed to the antioxidative property of licorice.

Diagnostic utility of CK19 and CD56 in the differentiation of thyroid papillary carcinoma from its mimics.

BACKGROUND: Papillary Thyroid Carcinoma (PTC) is the most common type of malignant thyroid tumors. The main diagnostic clue of PTC is the presence of its characteristic nuclear features. Yet, the focal presence of these features in other thyroid lesions causes a diagnostic dilemma. AIM: To evaluate Cytokeratin 19 (CK19) and CD56 immunostains as useful diagnostic markers in distinguishing papillary thyroid carcinoma from other mimicking thyroid lesions. METHODS: Eighty cases of different thyroid lesions were submitted for immunohistochemical staining of CK19 and CD56. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy were analyzed. RESULTS: CK19 was expressed in 87.8% and 21.2% of the PTC group and the non-papillary carcinoma (NPTC) group respectively, with significant difference (P<0.001). CD56 expression was lost with 81.8% of the PTC group. However, CK19 was negative in only 12.7% of the NPTC group, with significant difference (P<0.001). Comparing papillary carcinoma with papillary hyperplasia, CK19 was the most sensitive immunostain and CD56 was the most specific one, with better diagnostic accuracy in combining both immunostains. Co-expression of CK19/CD56 provided 100% sensitivity and 92% diagnostic accuracy in differentiating follicular variant of PTC from follicular adenoma. Comparing FVPTC with follicular carcinoma, sensitivity and diagnostic accuracy increased to 100% and 91.7% respectively. On distinction between papillary carcinoma (Hurthle cell variant) and Hurthle cell adenoma, sensitivity, specificity, and diagnostic accuracy were 100%, 75% and 83.3% respectively, with CK19/CD56 staining combination. Comparing papillary carcinoma on top of Hashimoto's thyroiditis with Hashimoto's thyroiditis, co-expression of both markers was associated with 100% specificity, as well as increase in PPV and diagnostic accuracy to 91%. CONCLUSION: The combined use of CK19 and CD56 is helpful in discriminating papillary thyroid carcinoma and its variants from other mimicking thyroid lesions.

Individual and combined effect of chlorpyrifos and cypermethrin on reproductive system of adult male albino rats.

Commercial mixtures of chlorpyrifos and cypermethrin pesticides are widely used to enhance the toxic effects of cypermethrin on target insects. So, the purpose of the current study was to evaluate the individual and combined toxic effects of chlorpyrifos (CPF) and cypermethrin (CYP) on reproductive system of adult male albino rats. Forty adult male albino rats were randomized into main four groups: group I (control group) included 16 rats, subdivided into negative and positive control; group II (eight rats) received chlorpyrifos 6.75 mg/kg b.w./orally∕daily); group III (eight rats) (received cypermethrin 12.5 mg/kg b.w./orally∕daily); and group IV (eight rats) (received chlorpyrifos and cypermethrin at the same previously mentioned doses). All treatments were given by oral gavage for 12 weeks. We found that single CPF and CYP exposures significantly have adverse effects on reproductive function of adult male albino rats manifested by reduced testicular weight, decreased sperm count, motility and viability, significantly increased percent of morphologically abnormal spermatozoa, and significant increments in sperm DNA fragmentation index (DFI) with respect to control group. Furthermore, serum follicle stimulating hormone, luteinizing hormone, and testosterone levels were decreased significantly compared to control group. This was accompanied with histopathological changes in the testis of rats such as necrosis, degeneration, decreasing number of spermatogenic cells in some seminiferous tubules, edema, congested blood vessels, and exudate in interstitial tissue of the testis. Notably, all these changes were exaggerated in rats treated concomitantly with chlorpyrifos and cypermethrin rendering the mixture more toxic than the additive effects of each compound and causing greater damage on the reproductive system of male albino rats than the individual pesticides.

Prognostic implications of epithelial to mesenchymal transition related proteins (E-cadherin, Snail) and hypoxia inducible factor 1α in endometrioid endometrial carcinoma.

The epithelial-mesenchymal transition (EMT) is an important step in the invasion and metastasis of cancer. E-cadherin downregulation, which is essentially controlled by EMT-mediated proteins such as Snail, is a main molecular feature of this process. Tumor hypoxia is one of the essential biological phenomena that are associated with the development and progression of various solid tumors. Recently, hypoxia and hypoxia-inducible factor 1α (HIF-1α) signaling pathway were identified to have an essential role in the regulation of EMT phenotype. The aim of the study was to evaluate the prognostic impact of EMT-related proteins (E-cadherin, Snail) and HIF-1α in endometrioid endometrial carcinoma (EEC) among Egyptian women. Immunohistochemical evaluation of E-cadherin, Snail, and HIF-1α expression was performed using 50 cases of EEC. The relationship between protein expression and clinicopathological features was investigated. The frequency of immunopositivity for E-cadherin, Snail, and HIF-1α in our cases of EEC was 82%, 28%, and 66%, respectively. Reduced E-cadherin and increased nuclear expression of Snail as well as HIF-1α were significantly associated with histopathologic grade, clinical stage myometrial invasion, and lymph node involvement. Statistical analysis showed a positive correlation between HIF-1α overexpression and Snail upregulation (τ= +0.252, P= .025); however, E-cadherin expression level was inversely correlated with enhanced Snail expression (τ= -0.450, P< .001) as well as with HIF-1α overexpression (τ= -0.439, P< .001). The overall survival and progression-free survival were inversely related to Snail immunoreactivity and positively related to E-cadherin expression. E-cadherin and Snail have a predictive value in EEC. In conclusion, the current study reveals that both Snail and HIF-1α expressions are significantly associated with poor prognosis in EEC; however, E-cadherin expression is considered a marker of good prognosis. E-cadherin and Snail expression has a predictive value in EEC management.

Diagnostic value of IMP3 and mesothelin in differentiating pancreatic ductal adenocarcinoma from chronic pancreatitis.

BACKGROUND: The discrimination between pancreatic ductal adenocarcinoma (PDA) and chronic pancreatitis may be confusing at both clinical and radiologic levels. So, the search for biomarkers able to distinguish both clinical conditions is of great interest. AIM: This study was undertaken to assess the value of insulin-like growth factor II mRNA binding protein 3 (IMP3) and mesothelin to differentiate PDA from non-neoplastic/reactive pancreatic duct epithelium. METHODS: Immunohistochemical staining for IMP3 and mesothelin was performed on 40 formalin-fixed, paraffin-embedded tissue sections of PDA, 20 biopsies of chronic pancreatitis and 10 normal pancreatic tissue obtained from tumor-free surgical margins. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy were calculated. RESULTS: IMP3 immunoreactivity was observed in 34 of 40 (85%) cases of PDA. The staining reaction was moderate to strong in 30 (75%) cases and diffuse in 26 (65%) cases. Eighteen of 20 (90%) biopsies of chronic pancreatitis were negative for IMP3, while the other two cases (10%) showed weak and focal IMP3 immunoreactivity. On the other hand, mesothelin demonstrated positive immunoreactivity in 30 of 40 (75%) cases of PDA. The staining reaction was moderate to strong in 24 (60%) cases and diffuse in 22 (55%) cases. Sixteen of 20 (80%) biopsies of chronic pancreatitis were negative for mesothelin, while weak and focal mesothelin staining was detected in the other 4 cases. All normal pancreatic tissues were negative for IMP3 and mesothelin expression. IMP3 showed higher sensitivity (85%) and specificity (90%) than mesothelin (75% and 80%, respectively). CONCLUSIONS: Our results showed that IMP3 immunostaining has a higher sensitivity and specificity than mesothelin for the diagnosis of PDA. IMP3 and mesothelin may be useful markers in distinguishing neoplastic from reactive lesions of the pancreas in instances where this is impossible by morphology alone in surgical pathology practice.